Patients and Methods

On-line supplementary material

Methods

Biochemical and hormonal parameters

Analysis of serum calcium and phosphate were performed by photometry (ADVIA 2400,

Siemens Healthcare Diagnostics, Frankfurt, Germany). Intact parathyroid hormone

(PTH) was determined by chemiluminescence immunoassay (CLIA), (Centaur, Siemens

Healthcare Diagnostics, Frankfurt, Germany) with an assay reference range of 1.3 to 7.6

pmol/L. Total 25-(OH)-vitamin D3 was measured by luminescence immunoassay (LIA),

(LIAISON, Dia Sorin, Dietzenbach, Germany). Serum bone-specific alkaline

phosphatase was measured by an enzyme-linked immunosorbent assay (ELISA),

(ACCESS, Beckmann Coulter, Krefeld, Germany). Osteocalcin was determined by

electrochemiluminescence immunoassay (ECLIA), (Modular, Roche Diagnostics,

Mannheim, Germany). Urinary desoxypyridinoline (DPD Crosslinks) were analyzed by

LIA (Immulite, Siemens Healthcare Diagnostics, Frankfurt, Germany), data are given per

mmol of creatinine. IGF-I and IGFBP-3 were analyzed by a commercially available

radioimmunoassay (Mediagnost, Reutlingen, Germany); z-score values were calculated

by use of gender- and age-specific reference values (Mediagnost, Reutlingen,

Germany). Plasma estradiol concentrations were measured by CLIA (Centaur,

Healthcare Diagnostics, Frankfurt, Germany). Plasma testosterone concentrations were

determined by radioimmunoassay, following extraction and chromatography, (Liquid

Szintillations Counter, Perkin-Elmer LAS, Rodgau-Jügesheim, Germany). 2

Supplementary Figure legends

Supplementary Figure 1: Total bone mineral density (BMD, z-score values) at the distal radius as a function of patient age. There was a weak significant inverse correlation (r = -

0.286; p=0.0348).

Supplementary Figure 1